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Preparation method of fast-growing frankia powder

A technology of Frank's bacteria and Frank's, which is applied in biochemical equipment and methods, methods of using spores, preparation of mutants, etc., to achieve the effects of strong stress resistance, fast growth rate and good inoculation effect

Inactive Publication Date: 2018-09-14
中科国思生物科技研究(广州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Frankia was discovered 100 years ago, but the success of artificial cultivation was first isolated from sweet fern in the United States in 1976, but it is a typical "slow-growing bacteria". It takes 13-15 hours for a reproduction cycle. It takes several months to get 1g of dried bacteria powder

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment example 1

[0028] Implementation case one: a kind of preparation method of fast-growing Frankia bacterium powder, comprises the following steps:

[0029] S1. Breeding Frankia strains: select 2-year-old seabuckthorn root nodules collected from June to August, process and cultivate them by root nodule sectioning method, and finally screen to obtain pure strains of Frankia species;

[0030] The root nodule slicing method comprises the following steps: cleaning the root nodules, putting them into 95% ethanol for disinfection for 5 minutes; then transferring them to 0.1% mercuric chloride for further disinfection and sterilization for 8 minutes; then thoroughly cleaning them with sterile water, Thin slices cut into 2 mm were inserted into QMOD medium and cultured at 28°C.

[0031] S2. Treat the pure strain of Frankia obtained in step S1 by ultraviolet-potassium chloride-diethyl sulfate compound mutagenesis method to obtain fast-growing Frankia;

[0032] S3. Inoculate the fast-growing Frankia...

Embodiment example 2

[0035] Implementation case two: a preparation method of fast-growing Frankia bacteria powder, comprising the steps of:

[0036] S1. Breeding Frankia strains: select 2-year-old seabuckthorn root nodules collected from June to August, process and cultivate them by shaking table culture method, and finally screen to obtain pure strains of Frankia species;

[0037] The shaker culture method includes the following steps: clean the root nodules, select robust and plump nodules, and infiltrate them with 75% ethanol in a vacuum for 15 minutes; Wash away residual ethanol with phosphate buffer; transfer to 0.1% mercuric chloride solution for vacuum infiltration and disinfection for 12 minutes; put it into a sterile mortar, add a small amount of quartz sand to make homogenate; filter through 400-mesh nylon mesh, and put the filtrate into In the QMOD medium, shake culture was carried out on a shaker at 28° C. and 160 r / min.

[0038] S2. The pure strain of Frankia obtained in step S1 is pro...

Embodiment example 3

[0042] Implementation case three: a preparation method of fast-growing Frankia bacteria powder, comprising the following steps:

[0043] S1. Breeding Frankia strains: select 2-year-old root nodules of seabuckthorn collected from June to August, treat and cultivate them by enzymatic treatment, and finally screen to obtain pure strains of Frankia species;

[0044] The enzyme treatment method comprises the following steps: cleaning the nodules at the root, and selecting robust and plump tumor flaps; after disinfecting the surface of the tumor flaps, placing them in a solution of 0.65mol / L of 0.5% cellulase and 2% pectinase The mannitol enzymatic hydrolysis solution was incubated at 25°C for 15 hours; then sliced, the skin layer was broken with a dissecting needle to release the bacteria, and then the bacteria liquid was inoculated on the plate, and streak culture was carried out at 28°C.

[0045] S2. Treat the pure strain of Frankia obtained in step S1 by ultraviolet-potassium ch...

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Abstract

The invention relates to a preparation method of fast-growing frankia powder. The method comprises the following steps: S1, breeding a frankia strain; S2, treating the pure frankia strain, obtained instep S1, by using a gene mutagenesis method to obtain a fast-growing frankia strain; S3, inoculating the fast-growing frankia strain, obtained in the step S2, into sterilized fermented raw materialsin a fermentation tank to form a bacterial culture material, and carrying out fermenting cultivation on the bacterial culture material to form hyphae; S4, further differentiating the hyphae, obtainedin the step S3, into spores, and eluting with sterile water to obtain bacterial liquid; adding sterile activated carbon into the bacterial liquid for adsorbing the spores, and separating the spore-containing activated carbon from the liquid after the spore adsorption is completed so as to obtain thallus taking the activated carbon as a carrier; S5, enabling the thallus, obtained in the step S4, tobe subjected to freeze-drying treatment by using a vacuum freeze-drying method so as to obtain the bacterial powder. The preparation method, provided by the invention, of the fast-growing frankia powder is good in inoculation effect, rapid in growth speed and suitable for large-scale industrial production.

Description

technical field [0001] The invention relates to the field of microbial fertilizer additive processing, in particular to a preparation method of fast-growing Frankia bacteria powder. Background technique [0002] Symbiotic nitrogen-fixing bacteria refer to a class of microorganisms that can form specific nitrogen-fixing tissue structures—root nodules—with the roots of host plants. They are symbiotic with each other. Plants provide photosynthetic products to microorganisms, which are needed for the growth and development of microorganisms to fix nitrogen, and microorganisms provide fixed nitrogen nutrients to plants. Both sides are mutually beneficial. Such nitrogen-fixing bacteria include Azotobacter rhizogenes and Frankia. According to measurements, rhizobia can fix about 13.3 kilograms of pure nitrogen per hectare per year, equivalent to about 130 kilograms of standardized fertilizers, and almost all of these nitrogen can be used by plants. [0003] Frankia is the only Ac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N3/00C12N15/01C12N1/02C05F11/08
CPCC05F11/08C12N1/02C12N1/20C12N3/00C12N15/01
Inventor 邵辉李彪
Owner 中科国思生物科技研究(广州)有限公司
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